目的 建立沉香HPLC指纹图谱分析方法,并结合醇溶性浸出物与沉香四醇的含量比较分析天然沉香与人工沉香,为科学评价沉香药材品质提供新方法。方法 按《中国药典》(2015年版)进行醇溶性浸出物和沉香四醇的含量测定;采用Platisil ODS 98色谱柱(4.6 mm×250 mm,5 μm),乙腈-0.1%甲酸溶液梯度洗脱,流速0.7 mL·min-1,检测波长252 nm,柱温31 ℃,建立沉香HPLC指纹图谱。使用《中药指纹图谱相似度评价系统(2004 A版)》软件建立共有模式,计算图谱相似度,共有峰相对峰面积进行主成分分析。结果 13批天然沉香的指纹图谱相似度为0.021~0.856,醇溶性浸出物含量为10.1%~31.0%,沉香四醇含量为0.04%~2.83%。11批人工沉香的指纹图谱相似度为0.079~0.453,醇溶性浸出物含量为10.3%~31.3%,沉香四醇含量为0.14%~1.02%。13批天然沉香和11批人工沉香分别各自聚为两组,对照药材聚在天然沉香组。天然沉香与人工沉香的指纹图谱相似度具有显著性差异(P<0.05),人工沉香指纹图谱相似度与醇溶性浸出物含量呈正相关(P<0.05)。结论 HPLC指纹图谱结合主成分分析可简便、快速地区分天然沉香与人工沉香,为全面评价沉香药材品质提供参考。
Abstract
OBJECTIVE To establish the HPLC fingerprint analysis method for agarwood with compare the natural agarwood and artificial agarwood combined with the contents of ethanol-soluble extraction (E%) and agarotetrol (A%). METHODS The E% and A% were determined according to the ChP(2015). The chromatographic fingerprints of agarwood were established on a Platisil ODS C18 column (4.6 mm×250 mm, 5 μm) with mobile phase consisting of acetonitrile-0.1% formic acid. Gradient elution was performed at a flow rate of 0.7 mL·min-1, the detection wave length was set at 252 nm, and the column temperature was maintained at 31 ℃. The similarity evaluation system of traditional Chinese medicine chromatographic fingerprint (2004A) was used to establish the common pattern and calculate the similarity of chromatograms. Principal component analysis (PCA) was used with multivariate statistical analysis software for relative peak area of common chromatographic peak. RESULTS The chromatographic fingerprint similarity of 13 batches of natural agarwood was 0.021-0.856, the E% was 10.1%-31.0%, and the A% was 0.04%-2.83%, respectively. The chromatographic fingerprint similarity of 11 batches of artificial agarwood ranged from 0.079 to 0.453, and the E% and A% were 10.3%-31.3% and 0.14%-1.02%, respectively. Thirteen batches of natural agarwood and 11 batches of artificial agarwwod were divided into two groups respectively, and the reference crude drug was gathered in the natural agarwood group. The similarity between natural and artificial agarwood chromatographic fingerprint was significantly different (P<0.05). Meanwhile, the similarity of artificial agarwood chromatographic fingerprint was positively correlated with the E% (P<0.05). CONCLUSION The chromatographic fingerprint analysis combined with multivariate statistical analysis can easily and quickly distinguish natural and artificial agarwood, which provides a reference for the comprehensive evaluation of the quality of agarwood.
关键词
沉香 /
高效液相色谱法 /
指纹图谱 /
主成分分析
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Key words
agarwood /
HPLC /
chromatographic fingerprint /
principal component analysis
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脚注
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基金
广东省科技计划项目资助(2017A020213014);华南应用微生物国家重点实验室开放基金项目资助(SKLAM002-2018);2017年广东药科大学国家级大学生创新创业训练计划项目资助(201710573001)
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